IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. confocal microscopy confocal microscopy immunostaining immunostaining Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay sequence tag sequence tag identification sequence tag Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions human-skov_3 Homo sapiens ovarian adenocarcinoma cell line skov_3 Human ovarian adenocarcinoma cell line Derived from the ascitic fluid from a 64 year old caucasian female with an ovarian tumour. Forms moderately well-differentiated adenocarcinoma consistent with ovarian primary cells. confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. pull down pull down western blot western blot Immuno blot Immuno blot Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. SIK2 Is a Centrosome Kinase Required for Bipolar Mitotic Spindle Formation that Provides a Potential Target for Therapy in Ovarian Cancer. Cancer Cell (1535-6108) 2010 imex curation Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk 21-Dec-2010: Contacted by JYOTI. orchard Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ahmed AA., Lu Z., Jennings NB., Etemadmoghadam D., Capalbo L., Jacamo RO., Barbosa-Morais N., Le XF., null null., Vivas-Mejia P., Lopez-Berestein G., Grandjean G., Bartholomeusz G., Liao W., Andreeff M., Bowtell D., Glover DM., Sood AK., Bast RC. Cancer - Interactions investigated in the context of cancer A kinase assay was performed in the presence of gamma-32P-ATP followed by autoradiography. The bands representing GST-C-Nap1-ct were cut and subjected to LC-MS analysis for protein identification and to detect sites of phosphorylation. Source of SIK2 taken from manufacturers website. Source of NAP1 inferred from sequence length and residue mapping. Assays with gamma-tubulin could not be captured as this could not be ascribed to a single gene product. ahmed.ahmed@obs-gyn.ox.ac.uk Accepted 2010-SEP-31 by ORCHARD. 21-Dec-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation sik2_human Serine/threonine-protein kinase SIK2 SIK2 Qin-induced kinase SNF1LK2 KIAA0781 QIK Serine/threonine-protein kinase SNF1-like kinase 2 Salt-inducible kinase 2 protein protein human Homo sapiens Human MVMADGPRHLQRGPVRVGFYDIEGTLGKGNFAVVKLGRHRITKTEVAIKIIDKSQLDAVNLEKIYREVQIMKMLDHPHIIKLYQVMETKSMLYLVTEYAKNGEIFDYLANHGRLNESEARRKFWQILSAVDYCHGRKIVHRDLKAENLLLDNNMNIKIADFGFGNFFKSGELLATWCGSPPYAAPEVFEGQQYEGPQLDIWSMGVVLYVLVCGALPFDGPTLPILRQRVLEGRFRIPYFMSEDCEHLIRRMLVLDPSKRLTIAQIKEHKWMLIEVPVQRPVLYPQEQENEPSIGEFNEQVLRLMHSLGIDQQKTIESLQNKSYNHFAAIYFLLVERLKSHRSSFPVEQRLDGRQRRPSTIAEQTVAKAQTVGLPVTMHSPNMRLLRSALLPQASNVEAFSFPASGCQAEAAFMEEECVDTPKVNGCLLDPVPPVLVRKGCQSLPSNMMETSIDEGLETEGEAEEDPAHAFEAFQSTRSGQRRHTLSEVTNQLVVMPGAGKIFSMNDSPSLDSVDSEYDMGSVQRDLNFLEDNPSLKDIMLANQPSPRMTSPFISLRPTNPAMQALSSQKREVHNRSPVSFREGRRASDTSLTQGIVAFRQHLQNLARTKGILELNKVQLLYEQIGPEADPNLAPAAPQLQDLASSCPQEEVSQQQESVSTLPASVHPQLSPRQSLETQYLQHRLQKPSLLSKAQNTCQLYCKEPPRSLEQQLQEHRLQQKRLFLQKQSQLQAYFNQMQIAESSYPQPSQQLPLPRQETPPPSQQAPPFSLTQPLSPVLEPSSEQMQYSPFLSQYQEMQLQPLPSTSGPRAAPPLPTQLQQQQPPPPPPPPPPRQPGAAPAPLQFSYQTCELPSAASPAPDYPTPCQYPVDGAQQSDLTGPDCPRSPGLQEAPSSYDPLALSELPGLFDCEMLDAVDPQHNGYVLVN 9D7F9BF81FD65CCF cp250_human Centrosome-associated protein CEP250 Centrosomal protein 2 Centrosomal Nek2-associated protein 1 CEP250 CEP2 CNAP1 250 kDa centrosomal protein protein protein human Homo sapiens Human METRSPGLNNMKPQSLQLVLEEQVLALQQQMAENQAASWRKLKNSQEAQQRQATLVRKLQAKVLQYRSWCQELEKRLEATGGPIPQRWENVEEPNLDELLVRLEEEQQRCESLAEVNTQLRLHMEKADVVNKALREDVEKLTVDWSRARDELMRKESQWQMEQEFFKGYLKGEHGRLLSLWREVVTFRRHFLEMKSATDRDLMELKAEHVRLSGSLLTCCLRLTVGAQSREPNGSGRMDGREPAQLLLLLAKTQELEKEAHERSQELIQLKSQGDLEKAELQDRVTELSALLTQSQKQNEDYEKMIKALRETVEILETNHTELMEHEASLSRNAQEEKLSLQQVIKDITQVMVEEGDNIAQGSGHENSLELDSSIFSQFDYQDADKALTLVRSVLTRRRQAVQDLRQQLAGCQEAVNLLQQQHDQWEEEGKALRQRLQKLTGERDTLAGQTVDLQGEVDSLSKERELLQKAREELRQQLEVLEQEAWRLRRVNVELQLQGDSAQGQKEEQQEELHLAVRERERLQEMLMGLEAKQSESLSELITLREALESSHLEGELLRQEQTEVTAALARAEQSIAELSSSENTLKTEVADLRAAAVKLSALNEALALDKVGLNQQLLQLEEENQSVCSRMEAAEQARNALQVDLAEAEKRREALWEKNTHLEAQLQKAEEAGAELQADLRDIQEEKEEIQKKLSESRHQQEAATTQLEQLHQEAKRQEEVLARAVQEKEALVREKAALEVRLQAVERDRQDLAEQLQGLSSAKELLESSLFEAQQQNSVIEVTKGQLEVQIQTVTQAKEVIQGEVRCLKLELDTERSQAEQERDAAARQLAQAEQEGKTALEQQKAAHEKEVNQLREKWEKERSWHQQELAKALESLEREKMELEMRLKEQQTEMEAIQAQREEERTQAESALCQMQLETEKERVSLLETLLQTQKELADASQQLERLRQDMKVQKLKEQETTGILQTQLQEAQRELKEAARQHRDDLAALQEESSSLLQDKMDLQKQVEDLKSQLVAQDDSQRLVEQEVQEKLRETQEYNRIQKELEREKASLTLSLMEKEQRLLVLQEADSIRQQELSALRQDMQEAQGEQKELSAQMELLRQEVKEKEADFLAQEAQLLEELEASHITEQQLRASLWAQEAKAAQLQLRLRSTESQLEALAAEQQPGNQAQAQAQLASLYSALQQALGSVCESRPELSGGGDSAPSVWGLEPDQNGARSLFKRGPLLTALSAEAVASALHKLHQDLWKTQQTRDVLRDQVQKLEERLTDTEAEKSQVHTELQDLQRQLSQNQEEKSKWEGKQNSLESELMELHETMASLQSRLRRAELQRMEAQGERELLQAAKENLTAQVEHLQAAVVEARAQASAAGILEEDLRTARSALKLKNEEVESERERAQALQEQGELKVAQGKALQENLALLTQTLAEREEEVETLRGQIQELEKQREMQKAALELLSLDLKKRNQEVDLQQEQIQELEKCRSVLEHLPMAVQEREQKLTVQREQIRELEKDRETQRNVLEHQLLELEKKDQMIESQRGQVQDLKKQLVTLECLALELEENHHKMECQQKLIKELEGQRETQRVALTHLTLDLEERSQELQAQSSQIHDLESHSTVLARELQERDQEVKSQREQIEELQRQKEHLTQDLERRDQELMLQKERIQVLEDQRTRQTKILEEDLEQIKLSLRERGRELTTQRQLMQERAEEGKGPSKAQRGSLEHMKLILRDKEKEVECQQEHIHELQELKDQLEQQLQGLHRKVGETSLLLSQREQEIVVLQQQLQEAREQGELKEQSLQSQLDEAQRALAQRDQELEALQQEQQQAQGQEERVKEKADALQGALEQAHMTLKERHGELQDHKEQARRLEEELAVEGRRVQALEEVLGDLRAESREQEKALLALQQQCAEQAQEHEVETRALQDSWLQAQAVLKERDQELEALRAESQSSRHQEEAARARAEALQEALGKAHAALQGKEQHLLEQAELSRSLEASTATLQASLDACQAHSRQLEEALRIQEGEIQDQDLRYQEDVQQLQQALAQRDEELRHQQEREQLLEKSLAQRVQENMIQEKQNLGQEREEEEIRGLHQSVRELQLTLAQKEQEILELRETQQRNNLEALPHSHKTSPMEEQSLKLDSLEPRLQRELERLQAALRQTEAREIEWREKAQDLALSLAQTKASVSSLQEVAMFLQASVLERDSEQQRLQDELELTRRALEKERLHSPGATSTAELGSRGEQGVQLGEVSGVEAEPSPDGMEKQSWRQRLEHLQQAVARLEIDRSRLQRHNVQLRSTLEQVERERRKLKREAMRAAQAGSLEISKATASSPTQQDGRGQKNSDAKCVAELQKEVVLLQAQLTLERKQKQDYITRSAQTSRELAGLHHSLSHSLLAVAQAPEATVLEAETRRLDESLTQSLTSPGPVLLHPSPSTTQAASR DIP protein Q9BV73 original sequence version: 129 BC2B8A36E07B8272 cetn3_human Centrin-3 CETN3 CEN3 protein protein human Homo sapiens Human MSLALRSELVVDKTKRKKRRELSEEQKQEIKDAFELFDTDKDEAIDYHELKVAMRALGFDVKKADVLKILKDYDREATGKITFEDFNEVVTDWILERDPHEEILKAFKLFDDDDSGKISLRNLRRVARELGENMSDEELRAMIEEFDKDGDGEINQEEFIAIMTGDI EF67724B2D6C2680 sik2-c_nap1 2 8 enzyme target enzyme target substrate substrate neutral component neutral component n terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 enzyme enzyme neutral component neutral component n terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation Fig 4D, 4E. Phosphorylation of enzyme targets; the predominant serine phosphorylation site of C-Nap1 is S2392 at the predicted consensus phosphorylation sequence and to a lesser extent S2394 (LHHSLSHSLL). In addition, a second site (S2234) at the fifth potential coiled coil region of C-Nap1 was confirmed. sik2-c_nap1-1 4 8 unspecified role unspecified role bait bait n terminus protein a tag protein a tag protein A protein A undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey physical association physical association Fig 4F. Immunoprecipitation using rabbit IgG-coated beads of protein A-tagged carboxy-terminal C-Nap1 but not protein A alone coprecipitated endogenous SIK2 (Figure 4F) indicating that direct binding between SIK2 and exogenously expressed carboxy-terminal part of C-Nap1 occurs in cells. sik2-c_nap1-2 6 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Fig 4G. Co-expression of the kinase mutant SIK2 did not result in phosphorylation of the coexpressed C-Nap1. It is not clear to me what the precipitating antibody used was - I have assummed it was anti-C_Nap1, though it does say "(IP) performed using the indicated antibodieS". (this makes marginally more sense than anything else...) According to Supplement it is full length c_NAp1. sik2-c_nap-1 1 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) colocalization colocalization Fig 4I. In addition, endogenous SIK2 partially colocalized with ectopically expressed C-Nap1 in the centrosome of SKOv3 cells (Figure S4I). sik2-centrin 5 9 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig S3D. The only centrin mentioned in the supplement (antibody to) is Centrin 3. I have entered 2 GO terms - dont know which or both is most appropriate, as the Centriole is part of the Centrosome. sik2-c_nap 3 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) colocalization colocalization Fig S4I. . In addition, endogenous SIK2 partially colocalized with ectopically expressed C-Nap1 in the centrosome of SKOv3 cells (Figure S4I).